scholarly journals Biotinylated single-chain variable fragment-based enzyme-linked immunosorbent assay for glycocholic acid

The Analyst ◽  
2018 ◽  
Vol 143 (9) ◽  
pp. 2057-2065 ◽  
Author(s):  
Xiping Cui ◽  
Natalia Vasylieva ◽  
Ding Shen ◽  
Bogdan Barnych ◽  
Jun Yang ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Competitive Elisa ◽  
Scfv Antibody ◽  
Enzyme Linked Immunosorbent Assay ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
Indirect Competitive Elisa ◽  
Glycocholic Acid

An indirect competitive ELISA was developed for GCA detection based on biotinylated scFv antibody.

Production and characterization of a single-chain variable fragment-alkaline phosphatase fusion protein for glycocholic acid detection in a one-step enzyme-linked immunosorbent assay

2018 ◽  
Vol 10 (22) ◽  
pp. 2629-2635 ◽  
Author(s):  
Xiping Cui ◽  
Qiyi He ◽  
Ding Shen ◽  
Zhengyun Jiang ◽  
Yingshan Chen ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Fusion Protein ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
Glycocholic Acid ◽  
One Step

One-step enzyme-linked immunosorbent assay for glycocholic acid based on single-chain variable fragment-alkaline phosphatase fusion protein.

scholarly journals Construction of a Single-Chain Variable-Fragment Antibody against the Superantigen Staphylococcal Enterotoxin B

2010 ◽  
Vol 76 (24) ◽  
pp. 8184-8191 ◽  
Author(s):  
Pawan Kumar Singh ◽  
Ranu Agrawal ◽  
Dev Vrat Kamboj ◽  
Garima Gupta ◽  
M. Boopathi ◽  
...  
Keyword(s):  
Phage Display ◽  
Staphylococcal Enterotoxin ◽  
Scfv Antibody ◽  
Affinity Constant ◽  
Enzyme Linked Immunosorbent Assay ◽  
Staphylococcal Enterotoxin B ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
High Affinity ◽  
Enterotoxin B

ABSTRACT Staphylococcal food poisoning (SFP) is one of the most prevalent causes of food-borne illness throughout the world. SFP is caused by 21 different types of staphylococcal enterotoxins produced by Staphylococcus aureus. Among these, staphylococcal enterotoxin B (SEB) is the most potent toxin and is a listed biological warfare (BW) agent. Therefore, development of immunological reagents for detection of SEB is of the utmost importance. High-affinity and specific monoclonal antibodies are being used for detection of SEB, but hybridoma clones tend to lose their antibody-secreting ability over time. This problem can be overcome by the use of recombinant antibodies produced in a bacterial system. In the present investigation, genes from a hybridoma clone encoding monoclonal antibody against SEB were immortalized using antibody phage display technology. A murine phage display library containing single-chain variable-fragment (ScFv) antibody genes was constructed in a pCANTAB 5E phagemid vector. Phage particles displaying ScFv were rescued by reinfection of helper phage followed by four rounds of biopanning for selection of SEB binding ScFv antibody fragments by using phage enzyme-linked immunosorbent assay (ELISA). Soluble SEB-ScFv antibodies were characterized from one of the clones showing high affinity for SEB. The anti-SEB ScFv antibody was highly specific, and its affinity constant was 3.16 nM as determined by surface plasmon resonance (SPR). These results demonstrate that the recombinant antibody constructed by immortalizing the antibody genes from a hybridoma clone is useful for immunodetection of SEB.

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